TW202241492A - Pharmaceutical composition of glp-1/glp-2 dual agonists - Google Patents

Abstract

The present invention relates to pharmaceutical compositions comprising particular preservatives.

Description

Pharmaceutical composition of GLP-1/GLP-2 dual agonist

The present invention relates to pharmaceutical compositions containing specific preservatives. The pharmaceutical compositions according to the invention are particularly stable and have an advantageous shelf life.

Peptides are an important part of the pharmaceutical industry. Although great progress has been made in the production of active pharmaceutical ingredients (APIs), the production of peptide-based pharmaceuticals remains a major challenge. The challenges associated with the development of peptide formulations are often overlooked or overlooked.

In general, a peptide is defined as a polypeptide of fewer than 50 amine residues and generally lacks an organized tertiary or globular structure. Some employ secondary structures, but these tend to be limited, such as a single-turn α-helix. While its smaller size makes it easier to deliver across biobarriers than larger proteins, its formulation can be problematic.

Some of the formulation challenges associated with peptides include, inter alia: chemical instability; adoption of multiple conformers; their tendency to self-associate; and complex physical instabilities such as gel formation, amyloid formation, and/or precipitation .

The most common challenge is chemical degradation of peptides and proteins through degradation mechanisms such as isomerization, racemization, hydrolysis, deamidation, and oxidation. The amino acid sequence of a given peptide defines the extent to which that peptide is susceptible to deamidation and/or oxidation reactions. The rate of oxidation of specific residues, such as the Met residue, correlates with the degree of solvent exposure. Since the peptide does not have a spherical structure that can chelate reactive groups, the side chains of almost all residues in the peptide are completely exposed to the solvent, enabling maximum contact with reactive oxygen species. Deamidation involves the hydrolysis of the amide side chains of amino acid residues, such as Asn and Gln. Furthermore, the high flexibility of peptide chains results in high deamidation rates compared to more complex proteins. However, it is important to note that the nature of the amino acid after deamidation (eg, the amino acid after Asn) also affects the deamidation rate. Peptides lacking steric hindrance and the ability to hydrogen bond to Asn side chains can accelerate the reaction even further. Typically, scientists must consider and test the reaction rates exhibited by Asn-Gly, Asn-Ala, Asn-Ser, and Asn-Asp amino acid combinations to ensure a stable pharmaceutical composition. Maximum control over hydrolysis reactions, including deamidation, is achieved through a stable and reliable pH and buffer system. However, such stable and reliable pH and buffering systems will be affected by the addition of additional excipients to the composition.

Excipients are added to pharmaceutical compositions to enhance or maintain active ingredient solubility (solubilizers) and/or stability (buffers, antioxidants, chelating agents, low temperature and lyoprotectants). Excipients are important in parenteral formulations in many cases to ensure safety (antimicrobial preservatives), minimize post-injection pain and irritation (tonicity agents) and to control or prolong drug delivery ( polymer). These are all examples of positive or synergistic interactions between excipients and drug products. However, any excipient added to the composition has the potential to produce negative effects, such as impaired peptide solubility, activity and/or chemical/physical stability, increased self-aggregation or fibrosis, which in turn can make drug administration undesirable. Safety.

Preservatives may be added to pharmaceutical compositions, such as where multiple aliquots are being used or drawn from a container containing multiple doses of the medicament, to kill microbial contaminants that may be introduced into the composition. Pharmaceutical compositions can be sealed and stored in sterile conditions without preservatives, but when using the container holding the composition, any accidental introduction of microorganisms may render the contents unsuitable for medical use. Therefore, efficient preservation of the medicinal contents is important, especially when the composition is stored in large volumes for several administrations. If containers holding larger volumes of unpreserved pharmaceutical compositions are used, the lack of preservatives can mean that most of the contents are discarded. Preservatives advantageously enable storage of the pharmaceutical composition at low temperatures (eg, refrigerated at about 5°C), such as for months or years, or at higher temperatures, such as room temperature, even after a portion of the composition has been used Store for shorter periods of time, such as days or weeks.

However, despite the advantages of using preservatives, the inclusion of preservatives in pharmaceutical compositions can be problematic because preservatives can interact adversely with other components of the composition, especially the active ingredient. Such interactions can lead to reduced preservation, or reduced or complete absence of the medical efficacy of the pharmaceutical composition. For example, preservatives may cause chemical instability of the activity. In the case of peptide activity, preservatives may participate in or promote degradation reactions, such as isomerization, racemization, hydrolysis, deamidation, or oxidation of the peptide, resulting in loss of the pharmacological activity of the peptide. Alternatively or additionally, preservatives may detrimentally affect the physical stability of the peptide activity, enhance aggregation of the peptide into inactive covalent oligomers and/or cause the peptide to precipitate out of solution. Not only does this loss of physical stability reduce the medical efficacy of the peptide, but the formation of particulate matter has practical and safety implications if the composition is delivered by injection.

Given the greater sequence variety of peptides and thus the different chemical structures, it is inherently unpredictable whether a given substance will act as an effective preservative for a particular therapeutic peptide composition without adversely affecting the peptide in the manner described above.

The present invention relates to a pharmaceutical composition comprising selected peptides disclosed in WO2018104561 (eg compound 18 of WO2018104561), which describes these compounds and their uses in detail.

The present application provides compositions comprising one or more dual GLP-1/GLP-2 agonists and one or more buffers containing preservatives. In some aspects, the composition is an isotonic parenteral pharmaceutical composition suitable for administration to a human subject.

GLP-1/GLP-2 dual agonists are peptides. Specific preservatives have been identified that can be used in compositions comprising specific GLP-1/GLP-2 dual agonist peptides and specific buffers without substantially affecting the chemical or physical stability of the peptides. It has been surprisingly found that a particular preservative has no effect on chemical stability at all peptide concentrations and has only a slight effect on the oligomerization of the peptide in phosphate buffer at higher peptide concentrations. Furthermore, it was found that when peptide compositions comprising m-cresol or phenol were stored at high temperature (40° C.) for a shorter period of time (2 weeks), counterintuitively, peptides oligomerized at higher concentrations of peptide less than at lower concentrations.

The compositions of the invention thus benefit from the advantages associated with preservatives, namely the prevention or reduction of microbial contamination.

The preservative effect of the compositions of the invention has particular advantages such that the compositions are provided in a multiple dose administration setting. The compositions of the invention may be provided in a device for sequentially administering therapeutic doses of the composition at time intervals over an extended period of time. Preservatives prevent the growth of microorganisms in the composition over this period of time while maintaining the chemical and physical stability of the peptide. This has the practical benefit that the device only needs to be loaded once, rather than having to prepare a new dose for each administration.

In one aspect, a pharmaceutical composition according to the invention is used or adapted for administration in a multi-dose device.

In some aspects, the invention provides a composition comprising: (a) one or more GLP-1/GLP-2 dual agonists comprising the general formula A: H[Aib]EG-X5-F-X7-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD (A), Wherein X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and at least one of X5 and X7 is T, wherein [Ψ] refers to the L or D lysine residue wherein the albumin binding moiety binds to the GLP-1/GLP-2 dual agonist, and Wherein the albumin binding moiety is [K([17-carboxy-heptadecyl]-isoGlu)]; (b) one or more preservatives, wherein the one or more preservatives comprise or are m-cresol and/or phenol; and (c) Phosphate buffer.

In some aspects, specific and specific compositions, such as isotonic parenteral compositions, are described in detail in the specification of the invention.

The chemical stability of the GLP-1/GLP-2 dual agonist at time point Y in any of the test compositions disclosed herein can be expressed as the GLP-1/GLP-2 dual agonist Relative purity X ^Y , and determined by measuring the absolute purity X' of the GLP-1/GLP-2 dual agonist at day zero (Day 0) for the GLP-1/GLP-2 dual agonist This is normalized to the absolute purity X ⁰ determined by HPLC at a given time point Y by identifying the purity of the peak corresponding to the GLP-1/GLP-2 dual agonist.

It was surprisingly found that the disclosed GLP-1/GLP-2 dual agonist in a composition comprising phosphate buffer and a preservative selected from m-cresol and phenol has good chemical stability at all peptide concentrations. Furthermore, it was found that when GLP-1/GLP-2 dual agonists were stored together with m-cresol or phenol at elevated temperature (40°C) for a short period of time (2 weeks), counterintuitively, at higher concentrations Fewer peptides oligomerized at higher concentrations than at lower concentrations.

The present invention also provides a composition according to the invention for: (i) increase intestinal mass, improve intestinal function, increase intestinal blood flow, or repair intestinal damage or dysfunction in individuals in need thereof; or (ii) Prevention or treatment of malabsorption, ulcers, short bowel syndrome, blind tube syndrome, inflammatory bowel disease, irritable bowel syndrome, pouchitis, celiac disease, tropical aphth, hypogammaglobulinemia aphthous ulcers in individuals in need , mucositis induced by chemotherapy or radiation therapy, diarrhea induced by chemotherapy or radiation therapy, low-grade inflammation, metabolic endotoxemia, necrotizing enterocolitis, primary biliary cirrhosis, hepatitis, fatty Gastrointestinal side effects of liver disease or inflammatory conditions; or (iii) reduces or inhibits weight gain, reduces gastric emptying or intestinal transit, reduces food intake, reduces appetite, or promotes weight loss in a subject in need thereof; or (iv) preventing or treating obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, inadequate glucose control, glucose tolerance, dyslipidemia, diabetes, prediabetes, metabolic syndrome, or hypertension.

The present invention further provides a method of preserving a composition comprising one or more GLP-1/GLP-2 dual agonists of the present invention and a phosphate buffer, wherein the method comprises adding one or more preservatives to the composition, Wherein the one or more preservatives comprise or are m-cresol and/or phenol.

The present invention further provides the use of a preservative in preserving a composition comprising one or more GLP-1/GLP-2 dual agonists of the present invention and a phosphate buffer, wherein the preservative comprises or is m-cresol and/or or phenol.

Compounds The compositions of the present invention comprise one or more GLP-1/GLP-2 dual agonists comprising the general formula A: H[Aib]EG-X5-F-X7-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD (A), Wherein X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and at least one of X5 and X7 is T, wherein [Ψ] is refers to the L or D lysine residue, wherein the albumin binding moiety binds to the GLP-1/GLP-2 dual agonist, and wherein the albumin binding moiety is [K([17-carboxy-heptadecyl ]-iso-Glu)].

In some aspects, one or more GLP-1/GLP-2 dual agonists comprising formula A have formula B: H[Aib]EG-X5-FT-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD (B), Wherein X5 is T or S and X28 is Q, E, A, H, Y, L, K, R or S, wherein [Ψ] refers to L or D lysine residues, wherein the albumin-binding moiety binds to GLP- 1/ GLP-2 dual agonist binding, and wherein the albumin binding moiety is [K([17-carboxy-heptadecyl]-isoGlu)].

In some aspects, one or more GLP-1/GLP-2 dual agonists comprising Formula A or B comprise the following sequence: H[Aib]EGSFTSELATILD[Ψ]QAARD FIAWLIQHKITD (SEQ ID NO: 1). In some aspects, the one or more GLP-1/GLP-2 dual agonists comprising Formula A or B consist of the sequence: H[Aib]EGSFTSELATILD[Ψ]QAARDFIAWLIQHKITD (SEQ ID NO: 1).

In some aspects, one or more GLP-1/GLP-2 dual agonists comprising the general formula A is: Hy-H[Aib]EGSFTSELATILD[K([17-Carboxyheptadecyl]-isoGlu )] QAARDFIAWLIQHKITD-OH (CPD1OH) or any pharmaceutically acceptable salt thereof. In some aspects, the pharmaceutically acceptable salt of CPD1OH is sodium salt, chloride salt or acetate salt, preferably chloride salt.

In some aspects, one or more GLP-1/GLP-2 dual agonists comprising the general formula A is: Hy-H[Aib]EGSFTSELATILD[K([17-Carboxyheptadecyl]-isoGlu )] QAARDFIAWLIQHKITD-NH ₂ (CPD1NH ₂ ) or any pharmaceutically acceptable salt thereof. In some aspects, the pharmaceutically acceptable salt of _CPD1NH2 is sodium salt, chloride salt or acetate salt, preferably chloride salt.

In a preferred aspect, the one or more GLP-1/GLP-2 dual agonists are CPD1OH or any pharmaceutically acceptable salt thereof, preferably its chloride salt.

As used herein, the term "GLP-1/GLP-2 dual agonist" refers to a peptide having activity at both the GLP-1 receptor and the GLP-2 receptor. A GLP-1/GLP-2 dual agonist comprising formula A or B may be a peptide of SEQ ID NO: 1 or a peptide in which one or more amino acids related to SEQ ID NO: 1 have been modified. Such agonists and/or peptides may further comprise one or more side chains that have been covalently attached to the GLP-1/GLP-2 dual agonist. The term "side chain" may also be referred to as "substituent".

As used herein, the term "salt" refers to an ionic compound that can be formed by the neutralization reaction of an acid and a base. Salts are composed of relevant numbers of cations (positively charged ions) and anions (negative ions) such that the product is charge neutral (no net charge). These component ions may be inorganic, such as chloride (Cl ⁻ ), or organic, such as acetate (CH ₃ CO ^{2 −} ); and may be monoatomic, such as fluoride (F ⁻ ), or polyatomic, Such as sulfate (SO ₄ ^{2 -} ).

As used herein, the term "pharmaceutically acceptable salt of CPD1" or "salt of CPD1" describes a salt of the compound comprising SEQ ID NO:1. As used herein, "Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecyl]-isoGlu)]QAARDFIAWLIQHKITD-OH. [acid]" identifies Hy-H[Aib]EGSFTSELATILD[K( The salt of [17-carboxy-heptadecyl]-isoGlu)]QAARDFIAWLIQHKITD-OH, wherein [acid] means that the compound is formed in a neutralization reaction, such as Hy-H[Aib]EGSFTSELATILD[K([17- The acid of the salt of carboxy-heptadecyl]-isoGlu)]QAARDFIAWLIQHKITD-OH. [HCl] will thus refer to the chloride salt.

As used herein, "pharmaceutically acceptable salt" refers to a salt that is safe and effective for use in mammals and possesses the desired biological activity. Pharmaceutically acceptable salts include salts of acidic or basic groups present in CPD1. For pharmaceutically acceptable salts, see Berge et al., 66 J. Pharm. Sci. 1-19 (1977), which is incorporated herein by reference. Table 1 - Selected GLP-1/GLP-2 dual agonists included in compositions of the invention SEQ ID CPD CPD form compound 1 1 1OH Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecyl]-isoGlu)]QAARDFIAWLIQHKITD-OH or a pharmaceutically acceptable salt thereof. 1 1 1NH ₂ Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecyl]-isoGlu)]QAARDFIAWLIQHKITD-NH ₂ or a pharmaceutically acceptable salt thereof.

The abbreviation CPD1 refers to any form of the compound comprising SEQ ID NO:1. However, CPD1OH only discloses compounds comprising SEQ ID NO: 1, wherein the compound is in its -OH form (free acid). CPD1NH2 form refers to the _-NH2 form (amidated form ₎ of the compound. Both CPD1OH and _CPD1NH2 can be converted into pharmaceutically acceptable salts to provide the drug substance in powder form. Table 2 - Amino acid sequences contained in one or more GLP-1/GLP-2 dual agonists of the invention SEQ ID sequence variable 1 H[Aib]EGSFTSELATILD[Ψ]QAARDFIAWLIQHKITD none 2 (Formula A) H[Aib]EG-X5-F-X7-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD X5 is T, S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S 3 (Formula B) H[Aib]EG-X5-FT-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD X5 is T, S; X28 is Q, E, A, H, Y, L, K, R or S

The "albumin-binding part" promotes the circulation of the GLP-1/GLP-2 dual agonist in the blood stream, and also has the effect of prolonging the action time of the GLP-1/GLP-2 dual agonist. The albumin binding moiety binds the GLP-1/GLP-2 dual agonist to albumin present in the blood, and because the GLP-1/GLP-2 dual agonist is only slowly released from albumin, the GLP-1/GLP -2 Prolongation of action of dual agonists. The term "albumin binding moiety" may also be referred to as a "side chain" or a "substituent".

As used herein, the term "natural amino acid" is an amino acid (common three-letter codes and one-letter codes in parentheses) selected from the group consisting of: glycine (Gly and G), proline Acids (Pro and P), Alanine (Ala and A), Valine (Val and V), Leucine (Leu and L), Isoleucine (Ile and I), Methionine (Met and M), cysteine (Cys and C), phenylalanine (Phe and F), tyrosine (Tyr and Y), tryptophan (Trp and W), histidine (His and H), lysine acid (Lys and K), arginine (Arg and R), glutamine (Gln and Q), asparagine (Asn and N), glutamic acid (Glu and E), aspartic acid ( Asp and D), serine (Ser and S) and threonine (Thr and T). If not otherwise indicated, amino acids indicated by their one-letter codes in uppercase letters indicate the L-isomer, whereas if amino acids are indicated by lowercase letters, the amino acids are indicated in their D-form, e.g. K (ie L-lysine), k (ie D-lysine) forms are used/applied.

The abbreviation "Hy-" in relation to compounds disclosed herein refers to hydrogen. The chosen abbreviation is indicated as "Hy" to avoid confusion of hydrogen with histidine (H) at the beginning of the sequence.

Throughout this specification and claims, the generally accepted three-letter codes for other "α-amino acids" are used, such as sarcosine (Sar), norleucine (NIe), α-aminoisobutyl acid (Aib), 2,3-diaminopropionic acid (Dap), 2,4-diaminobutyric acid (Dab) and 2,5-diaminopentanoic acid (ornithine; Orn). When used in a formula or sequence in this specification, especially when a single-letter code is used to represent the remainder of the formula or sequence, such other α-amino acids may be shown within square brackets "[ ]" (eg, " [Aib]").

Compound Concentration In some aspects, compositions of the invention, such as isotonic parenteral pharmaceutical compositions of the invention, comprise at least about 1 mg/mL of a GLP-1/GLP-2 dual agonist, such as at least about 2 mg/mL GLP-1/GLP-2 dual agonist. In some aspects, a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, comprises at least about 1 mg/mL to about 33 mg/mL of a GLP-1/GLP-2 dual agonist , such as at least about 2 mg/mL to about 33 mg/mL of a GLP-1/GLP-2 dual agonist. In some aspects, compositions of the invention, such as isotonic parenteral pharmaceutical compositions of the invention, comprise at least about 1 mg/mL to about 25 mg/mL of a GLP-1/GLP-2 dual agonist , such as at least about 2 mg/mL to about 25 mg/mL of a GLP-1/GLP-2 dual agonist, such as at least about 4 mg/mL to about 25 mg/mL of a GLP-1/GLP-2 dual agonist A potent agent, such as a GLP-1/GLP-2 dual agonist of at least about 6 mg/mL to about 25 mg/mL, such as a GLP-1/GLP-2 of at least about 8 mg/mL to about 25 mg/mL Dual agonists, such as at least about 10 mg/mL to about 25 mg/mL of a GLP-1/GLP-2 dual agonist.

In some aspects, a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, comprises about 1 mg/mL of a GLP-1/GLP-2 dual agonist. In some aspects, compositions of the invention, such as isotonic parenteral pharmaceutical compositions of the invention, comprise about 2 mg/mL of a GLP-1/GLP-2 dual agonist. In some aspects, a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, comprises about 4 mg/mL of a GLP-1/GLP-2 dual agonist. In some aspects, a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, comprises about 6 mg/mL of a GLP-1/GLP-2 dual agonist. In some aspects, a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, comprises about 8 mg/mL of a GLP-1/GLP-2 dual agonist. In some aspects, a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, comprises about 10 mg/mL of a GLP-1/GLP-2 dual agonist. In some aspects, a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, comprises about 15 mg/mL of a GLP-1/GLP-2 dual agonist. In some aspects, compositions of the invention, such as isotonic parenteral pharmaceutical compositions of the invention, comprise about 20 mg/mL of a GLP-1/GLP-2 dual agonist. In some aspects, a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, comprises about 25 mg/mL of a dual GLP-1/GLP-2 agonist. In some aspects, a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, comprises about 33 mg/mL of a GLP-1/GLP-2 dual agonist.

Preferably, the compositions of the present invention comprise from about 6 mg/mL to about 25 mg/mL GLP-1/GLP-2 dual agonist or from about 2 mg/mL to about 10 mg/mL GLP-1/GLP-2 2 dual agonists. Optimally, the compositions of the invention comprise about 15 mg/mL of the GLP-1/GLP-2 dual agonist.

Synthesis of dual agonists Preferably, the dual agonists of the invention are synthesized by means of solid phase or solution phase peptide synthesis methods. In this context, reference may be made to WO 98/11125, and especially to Fields, G.B. et al., 2002, "Principles and practice of solid-phase peptide synthesis". In: Synthetic Peptides (2nd Edition) and in the Examples herein . According to the present invention, the dual agonists of the present invention can be synthesized or produced in a variety of ways, including, for example, methods comprising: (a) synthesizing the dual agonist by means of solid-phase or liquid-phase peptide synthesis methods and recovering the synthesized dual agonist thus obtained; or (b) expressing a precursor peptide sequence from a nucleic acid construct encoding the precursor peptide, recovering the expressed product and modifying the precursor peptide to produce a compound of the invention.

The precursor peptide can be modified by introducing one or more non-proteinaceous amino acids, such as Aib, Orn, Dap or Dab; introducing an albumin binding moiety, or introducing an appropriate terminal group -OH or _-NH2 , etc.

Expression is usually from nucleic acid encoding the precursor peptide, which can be in a cell or a cell-free expression system comprising such nucleic acid.

Preservatives Compositions of the invention, such as the isotonic parenteral pharmaceutical compositions of the invention, comprise one or more preservatives. In some aspects, the one or more preservatives include or are m-cresol and/or phenol. In some aspects, the preservative is m-cresol or phenol. The compositions of the present invention, such as the isotonic parenteral pharmaceutical composition of the present invention, contain a preservative, wherein the preservative comprises or is m-cresol and/or phenol. In some aspects, compositions of the invention, such as the isotonic parenteral pharmaceutical compositions of the invention, comprise a preservative, wherein the preservative comprises or is m-cresol or phenol. In some aspects, a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, comprises two preservatives, wherein the preservatives comprise or are m-cresol or phenol.

m-cresol In some aspects, compositions of the invention, such as isotonic parenteral pharmaceutical compositions of the invention, comprise one or more preservatives, wherein one or more preservatives comprise m-cresol. In some aspects, the one or more preservatives are m-cresol. In some aspects, compositions of the invention, such as the isotonic parenteral pharmaceutical compositions of the invention, comprise a m-cresol preservative. In some aspects, compositions of the invention, such as the isotonic parenteral pharmaceutical compositions of the invention, comprise m-cresol.

。 Meta-cresol is an organic compound also known as m-cresol, 3-cresol, 3-cresol, 3-methylhydroxybenzene, 3-hydroxytoluene, or 1-hydroxy-3-toluene. m-cresol has the chemical formula CH ₃ C ₆ H ₄ (OH) and the following structural formula:

.

In some aspects, m-cresol is present in the compositions of the invention at a concentration of about 1.15 mg/mL to about 5.15 mg/mL. In some aspects, m-cresol is present in the compositions of the invention at a concentration of about 1.15 mg/mL. In some aspects, m-cresol is present in the compositions of the invention at a concentration of about 5.15 mg/mL. Preferably, m-cresol is present in the compositions of the present invention at a concentration of about 3.15 mg/mL.

Phenol In some aspects, compositions of the invention, such as isotonic parenteral pharmaceutical compositions of the invention, comprise one or more preservatives, wherein the one or more preservatives comprise phenol. In some aspects, the one or more preservatives are phenol. In some aspects, compositions of the invention, such as the isotonic parenteral pharmaceutical compositions of the invention, comprise a preservative, which is phenol. In some aspects, compositions of the invention, such as the isotonic parenteral pharmaceutical compositions of the invention, comprise phenol.

。 Phenol is an organic compound also known as hydroxybenzene. Phenol has the chemical _{formula C6H5OH} and the following structural formula:

.

In some aspects, phenol is present in the compositions of the invention at a concentration of about 2.5 mg/mL to about 8.5 mg/mL. In some aspects, phenol is present in the compositions of the invention at a concentration of about 2.5 mg/mL. In some aspects, phenol is present in the compositions of the invention at a concentration of about 8.5 mg/mL. Preferably, phenol is present in the compositions of the invention at a concentration of about 5.5 mg/mL.

Phosphate buffer and pH Compositions of the invention, such as the isotonic parenteral pharmaceutical compositions of the invention, comprise phosphate buffered saline.

In some aspects, the phosphate buffer is present in the composition at a final concentration of about 5 mM to about 50 mM, such as about 5 mM to about 40 mM, such as about 5 mM to about 30 mM, such as isotonic Enteric medicinal composition. Preferably, phosphate buffer is present in the composition at a final concentration of from about 5 mM to about 20 mM. In some aspects, phosphate buffer is present in the composition at a final concentration of about 5 mM. In some aspects, phosphate buffer is present in the composition at a final concentration of about 50 mM. Optimally, phosphate buffer is present in the composition at a final concentration of about 20 mM.

In some aspects, the phosphate buffer is sodium phosphate buffer. In some aspects, the phosphate buffer is disodium phosphate (Na ₂ HPO ₄ ) or sodium dihydrogen phosphate (NaH ₂ PO ₄ ), or a combination thereof.

In one aspect, disodium phosphate is present in the composition at a final concentration of about 15 mM to about 19 mM, preferably between 18 mM and 19 mM.

In one aspect, sodium dihydrogen phosphate is present in the isotonic parenteral pharmaceutical composition at a final concentration of about 1 mM to about 3 mM, preferably between 1 mM and 2 mM.

In one aspect, the final concentration of the disodium phosphate and sodium dihydrogen phosphate buffer components together is from about 5 mM to about 50 mM, preferably from about 10 mM to about 40 mM, more preferably from about 15 mM to about 30 mM . In an optimal aspect, the final concentration of the disodium phosphate and sodium dihydrogen phosphate buffer components together is about 20 mM.

In some aspects, the pH of the compositions of the invention, such as isotonic parenteral pharmaceutical compositions, is between about pH 6.0 and about pH 8.5, for example between about pH 6.0 and about pH 8.4, at about pH 6.0 and about pH 8.3, between about pH 6.0 and about pH 8.2, between about pH 6.0 and about pH 8.1 or between about pH 6.0 and about pH 8.0. In some aspects, the pH is a pH of about pH 6.5 to about pH 8.5. In one aspect, the pH is preferably between about pH 7.0 and about pH 8.0. In some aspects, the pH of compositions of the invention, such as isotonic parenteral pharmaceutical compositions, is between about pH 7.0 to about pH 8.0. In some aspects, the pH of compositions of the invention (such as isotonic parenteral pharmaceutical compositions) is about pH 7.0. In some aspects, compositions of the invention (such as isotonic parenteral pharmaceutical compositions ) is about pH 8.0. In some aspects, the pH of the compositions of the present invention is about pH 8.2. In some aspects, the pH of the compositions of the present invention is about pH 6.0. In some aspects In some aspects, the pH of the compositions of the present invention is between about pH 7.0 and about pH 8.2, preferably about pH 7.5 or about pH 8.2. In some aspects, the pH of the compositions of the present invention is between about pH 7.0 and about pH 8.2. Between about pH 8.2, preferably about pH 7.6 or about 8.0. In some aspects, the pH of the composition of the present invention is between about pH 7.0 and about pH 8.2, preferably about pH 7.6 or about pH 7.7. In some aspects, the pH of the compositions of the present invention is between about pH 7.0 and about pH 8.2, preferably about pH 7.6. In some aspects, the pH of the compositions of the present invention is between about pH 7.0 and about pH 8.2. 8.2, preferably about pH 8.0. In some aspects, the pH of the compositions of the present invention is between about pH 7.0 and about pH 8.2, preferably about pH 7.0. In a preferred aspect , pH is about 8.0.

In some aspects, in the compositions of the invention, the pH is adjusted with NaOH or HCl as needed.

Tonicity and Tonicity Agents In some aspects, the compositions of the invention are isotonic parenteral pharmaceutical compositions.

In some aspects, compositions of the invention comprising one or more GLP-1/GLP-2 dual agonists comprising Formula A or B are isotonic. In some aspects, compositions of the invention comprising one or more GLP-1/GLP-2 dual agonists comprising SEQ ID NO: 1 are isotonic.

In some embodiments, the compositions of the present invention have an osmolality of about 300±120 mOsmol/kg. In some embodiments, compositions of the present invention have an osmolality of about 290±70 mOsmol/kg. In some embodiments, the compositions of the present invention have an osmolality of from about 230 mOsmol/kg to about 370 mOsmol/kg. In some embodiments, the compositions of the present invention have an osmolality of from about 280 mOsmol/kg to about 320 mOsmol/kg. In some embodiments, the compositions of the present invention have an osmolality of from about 290 mOsmol/kg to about 320 mOsmol/kg.

In some aspects, compositions of the invention, such as isotonic parenteral pharmaceutical compositions of the invention, comprise one or more tonicity agents.

The term "tonicity agent" refers to an excipient which is added to the composition according to the invention in order to achieve isotonicity in relation to bodily fluids. A range of ionic and nonionic tonicity agents are used in pharmaceutical compositions. Nonionic tonicity agents may be selected from dextrose, propylene glycol, glycerol, mannitol (such as D-mannitol), and sorbitol. Ionic tonicity agents may include alkali metal or alkaline earth metal halides such as CaCl ₂ , KBr, KCl, LiCl, NaI, NaBr, NaCl, Na ₂ SO ₄ . In one aspect, the tonicity agent may be selected from mannitol, NaCl, and propylene glycol.

An "ionic compound" is two or more ions held together by attractive forces. An example of an ionic compound is table salt. It consists of positive sodium ions and negative chloride ions. It has high melting and boiling points and is hard or brittle. It is also soluble in water. A "nonionic compound" is defined as a compound in which the chemical linkages are nonionic. They generally have chemical bonds that share electron density.

In some aspects, the one or more tonicity agents comprise or are mannitol. Preferably, the one or more tonicity agents are D-mannitol. In some aspects, mannitol (such as D-mannitol) is present in the present invention at a concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, more preferably about 190 mM to about 240 mM. In the composition of the invention. In some aspects, mannitol, such as D-mannitol, is present in the compositions of the invention at about 230 mM.

In some aspects, the one or more tonicity agents comprise or are NaCl. In some aspects, NaCl is present in the compositions of the invention at a concentration of about 50 mM to about 450 mM, preferably about 65 mM to about 165 mM. Preferably, NaCl is present at a concentration of about 125 mM.

As used herein, the term "isotonic" refers to the tension associated with a body fluid at the site of injection (ie, intravenous or subcutaneous). Thus, the term "isotonic" is used to describe a pharmaceutical composition having the same tonicity as a body fluid, such as red blood cells and/or plasma, at the site of injection. A composition having an osmolality of about 300 mOsmol/kg, such as about 280-320 mOsmol/kg or about 290-320 mOsmol/kg is considered isotonic.

Isotonicity is critical for parenteral pharmaceutical compositions, since "hypotonic" solutions cause cells to swell, while "hypertonic" solutions cause cells to contract. While tonicity is related to osmolality, tonicity also takes into account the ability of a solute to cross a cell membrane.

Compositions of the Invention In some aspects, compositions of the invention, such as the isotonic parenteral pharmaceutical compositions of the invention, comprise a solvent. In some aspects, the solvent is water.

In some aspects, compositions of the invention, such as the isotonic parenteral pharmaceutical compositions of the invention, include tonicity agents and solvents.

In some aspects, the composition of the present invention (such as the isotonic parenteral pharmaceutical composition of the present invention) comprises a GLP-1/GLP-2 dual agonist comprising the amino acid sequence of formula A, m-cresol and phosphate buffered saline.

In some aspects, compositions of the invention (such as the isotonic parenteral pharmaceutical composition of the invention) comprise a GLP-1/GLP-2 dual agonist comprising the amino acid sequence of formula A, phenol, and phosphoric acid salt buffer.

In some aspects, the composition of the present invention (such as the isotonic parenteral pharmaceutical composition of the present invention) comprises a GLP-1/GLP-2 dual agonist comprising the amino acid sequence of formula A, m-cresol , phosphate buffer, and mannitol, such as D-mannitol.

In some aspects, compositions of the invention (such as the isotonic parenteral pharmaceutical composition of the invention) comprise a GLP-1/GLP-2 dual agonist comprising the amino acid sequence of formula A, phenol, phosphoric acid Saline buffer and mannitol, such as D-mannitol.

In some aspects, in the composition of the invention (such as the isotonic parenteral pharmaceutical composition of the invention), the GLP-1/GLP-2 dual agonist comprises the amino acid sequence of formula A, a preservative Cresol at a final concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, or a preservative at a final concentration of about 2.5 mg/mL to about 8.5 mg/mL, preferably about 8.5 mg/mL 5.5 mg/mL of phenol, and the final concentration of phosphate buffer is about 5 mM to about 50 MM, preferably about 20 mM.

In some aspects, in the composition of the invention (such as the isotonic parenteral pharmaceutical composition of the invention), the GLP-1/GLP-2 dual agonist comprises the amino acid sequence of formula A, a preservative Cresol at a final concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, and a final concentration of phosphate buffer of about 5 mM to about 50 mM, preferably about 20 mM .

In some aspects, in the composition of the invention (such as the isotonic parenteral pharmaceutical composition of the invention), the GLP-1/GLP-2 dual agonist comprises the amino acid sequence of formula A, a preservative It is phenol at a final concentration of about 2.5 mg/mL to about 8.5 mg/mL, preferably about 5.5 mg/mL, and the final concentration of phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM.

In some aspects, in the composition of the invention (such as the isotonic parenteral pharmaceutical composition of the invention), the GLP-1/GLP-2 dual agonist comprises the amino acid sequence of formula A and is expressed in about 1 mg/mL to about 33 mg/mL, preferably about 1 mg/mL to about 25 mg/mL, preferably about 6 mg/mL to about 25 mg/mL, preservative at a final concentration of about 1.15 mg/mL mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL cresol, or the preservative is phenol at a final concentration of about 2.5 mg/mL to about 8.5 mg/mL, preferably about 5.5 mg/mL , and the final concentration of the phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM.

In some aspects, in the composition of the invention (such as the isotonic parenteral pharmaceutical composition of the invention), the GLP-1/GLP-2 dual agonist comprises the amino acid sequence of formula A and is expressed in about 1 mg/mL to about 33 mg/mL, preferably about 1 mg/mL to about 25 mg/mL, preferably about 6 mg/mL to about 25 mg/mL, preservative at a final concentration of about 1.15 mg/mL mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, and the final concentration of phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM.

In some aspects, in the composition of the invention (such as the isotonic parenteral pharmaceutical composition of the invention), the GLP-1/GLP-2 dual agonist comprises the amino acid sequence of formula A and is expressed in about 1 mg/mL to about 33 mg/mL, preferably about 1 mg/mL to about 25 mg/mL, preferably about 6 mg/mL to about 25 mg/mL, preservative at a final concentration of about 2.5 mg/mL to about 8.5 mg/mL, preferably about 5.5 mg/mL of phenol, and the final concentration of the phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM.

In some aspects, in the composition of the invention (such as the isotonic parenteral pharmaceutical composition of the invention), the GLP-1/GLP-2 dual agonist comprises the amino acid sequence of formula A and is expressed in about 1 mg/mL to about 33 mg/mL, preferably about 1 mg/mL to about 25 mg/mL, preferably about 6 mg/mL to about 25 mg/mL, more preferably about 25 mg/mL, The preservative is cresol at a final concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably between about 3.15 mg/mL, or the preservative is at a final concentration of about 2.5 mg/mL to about 8.5 mg/mL, more preferably Preferably about 5.5 mg/mL of phenol, and the final concentration of the phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM. The composition preferably also comprises mannitol, such as D-mannitol, at a concentration of from about 130 mM to about 330 mM, preferably from about 150 mM to about 300 mM, preferably from about 190 mM to about 240 mM, preferably about 230 mM sugar alcohol.

In some aspects, in the composition of the invention (such as the isotonic parenteral pharmaceutical composition of the invention), the GLP-1/GLP-2 dual agonist comprises the amino acid sequence of formula A and is expressed in about 1 mg/mL to about 33 mg/mL, preferably about 1 mg/mL to about 25 mg/mL, preferably about 6 mg/mL to about 25 mg/mL, more preferably about 25 mg/mL, The preservative is cresol at a final concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, and the final concentration of the phosphate buffer is about 5 mM to about 50 mM, preferably about 5 mM to about 50 mM. 20 mM. The composition preferably also comprises mannitol, such as D-mannitol, at a concentration of from about 130 mM to about 330 mM, preferably from about 150 mM to about 300 mM, preferably from about 190 mM to about 240 mM, preferably about 230 mM sugar alcohol.

In some aspects, in the composition of the invention (such as the isotonic parenteral pharmaceutical composition of the invention), the GLP-1/GLP-2 dual agonist comprises the amino acid sequence of formula A and is expressed in about 1 mg/mL to about 33 mg/mL, preferably about 1 mg/mL to about 25 mg/mL, preferably about 6 mg/mL to about 25 mg/mL, more preferably about 25 mg/mL, The preservative is phenol at a final concentration of about 2.5 mg/mL to about 8.5 mg/mL, preferably about 5.5 mg/mL, and the final concentration of phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM. The composition preferably also comprises mannitol, such as D-mannitol, at a concentration of from about 130 mM to about 330 mM, preferably from about 150 mM to about 300 mM, preferably from about 190 mM to about 240 mM, preferably about 230 mM sugar alcohol.

In some aspects, in a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, the dual GLP-1/GLP-2 agonist is CPD1OH or a pharmaceutically acceptable salt thereof , the final concentration is preferably from about 1 mg/mL to about 33 mg/mL, preferably from about 1 mg/mL to about 25 mg/mL, preferably from about 2 mg/mL to about 25 mg/mL, more preferably about 6 mg/mL to about 25 mg/mL, the preservative is cresol at a final concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, or the preservative is about 2.5 mg final concentration /mL to about 8.5 mg/mL, preferably about 5.5 mg/mL of phenol, and the final concentration of the phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM.

In some aspects, in a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, the dual GLP-1/GLP-2 agonist is CPD1OH or a pharmaceutically acceptable salt thereof , the final concentration is preferably from about 1 mg/mL to about 33 mg/mL, preferably from about 1 mg/mL to about 25 mg/mL, preferably from about 2 mg/mL to about 25 mg/mL, more preferably about 6 mg/mL to about 25 mg/mL, the preservative is cresol at a final concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, and the final concentration of the phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM.

In some aspects, in a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, the dual GLP-1/GLP-2 agonist is CPD1OH or a pharmaceutically acceptable salt thereof , the final concentration is preferably from about 1 mg/mL to about 33 mg/mL, preferably from about 1 mg/mL to about 25 mg/mL, preferably from about 2 mg/mL to about 25 mg/mL, more preferably about 6 mg/mL to about 25 mg/mL, the preservative is phenol at a final concentration of about 2.5 mg/mL to about 8.5 mg/mL, preferably about 5.5 mg/mL, and the final concentration of phosphate buffer is about 5 mM To about 50 mM, preferably about 20 mM.

In some aspects, in a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, the GLP-1/GLP-2 dual agonist is CPD1OH or a pharmaceutically acceptable salt thereof, The preservative is cresol at a final concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, and the final concentration of the phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM, and the tonicity agent is mannitol, such as D-mannitol, at a final concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably About 230 mM.

In some aspects, in a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, the dual GLP-1/GLP-2 agonist is CPD1OH or a pharmaceutically acceptable salt thereof , the preservative is phenol at a final concentration of about 2.5 mg/mL to about 8.5 mg/mL, preferably about 5.5 mg/mL, and the final concentration of the phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM , and the tonicity agent is mannitol, such as D-mannitol, the final concentration is about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

In some aspects, in a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, the dual GLP-1/GLP-2 agonist is CPD1OH or a pharmaceutically acceptable salt thereof , the final concentration is preferably from about 1 mg/mL to about 33 mg/mL, preferably from about 1 mg/mL to about 25 mg/mL, preferably from about 2 mg/mL to about 25 mg/mL, more preferably about 6 mg/mL to about 25 mg/mL, the preservative is cresol at a final concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, or the preservative is about 2.5 mg final concentration /mL to about 8.5 mg/mL, preferably about 5.5 mg/mL of phenol, the final concentration of phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM, and the tonicity agent is mannitol, such as D-mannitol, the final concentration is about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM.

In some aspects, in a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, the dual GLP-1/GLP-2 agonist is CPD1OH or a pharmaceutically acceptable salt thereof , the final concentration is preferably from about 1 mg/mL to about 33 mg/mL, preferably from about 1 mg/mL to about 25 mg/mL, preferably from about 2 mg/mL to about 25 mg/mL, more preferably about 6 mg/mL to about 25 mg/mL, the preservative is cresol at a final concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, and the final concentration of phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM, and the tonicity agent is mannitol, such as D-mannitol, at a final concentration of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably About 190 mM to about 240 mM, preferably about 230 mM.

In some aspects, in a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, the dual GLP-1/GLP-2 agonist is CPD1OH or a pharmaceutically acceptable salt thereof , the final concentration is preferably from about 1 mg/mL to about 33 mg/mL, preferably from about 1 mg/mL to about 25 mg/mL, preferably from about 2 mg/mL to about 25 mg/mL, more preferably about 6 mg/mL to about 25 mg/mL, the preservative is phenol at a final concentration of about 2.5 mg/mL to about 8.5 mg/mL, preferably about 5.5 mg/mL, and the final concentration of the phosphate buffer is about 5 mM to About 50 mM, preferably about 20 mM, and the tonicity agent is mannitol, such as D-mannitol, the final concentration is about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM mM to about 240 mM, preferably about 230 mM.

In some aspects, in a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, the dual GLP-1/GLP-2 agonist is CPD1OH or a pharmaceutically acceptable salt thereof , the preservative is cresol at a final concentration of about 3.15 mg/mL, the phosphate buffer is at a final concentration of about 20 mM, and the tonicity agent is mannitol, such as D-mannitol, at a final concentration of about 230 mM.

In some aspects, in a composition of the invention, such as an isotonic parenteral pharmaceutical composition of the invention, the dual GLP-1/GLP-2 agonist is CPD1OH or a pharmaceutically acceptable salt thereof , the preservative is phenol at a final concentration of about 5.5 mg/mL, the phosphate buffer is at a final concentration of about 20 mM, and the tonicity agent is mannitol, such as D-mannitol, at a final concentration of about 230 mM.

Indications In some aspects, the pharmaceutical compositions of the invention are administered to human subjects in need of preventing or treating intestinal injury and dysfunction, regulating body weight, and preventing or treating metabolic dysfunction.

In some aspects, a pharmaceutical composition of the invention is administered to a human subject in need of prevention or treatment of malabsorption, ulcers (e.g., gastric ulcer, Zollinger-Ellison Syndrome), drug-induced ulcers, and Ulcers associated with infection or other pathogens), short bowel syndrome, dead-leg syndrome, inflammatory bowel disease (Crohns disease and ulcerative colitis), irritable bowel syndrome (IBS), pouchitis, celiac disease (e.g. caused by gluten-induced enteropathy or celiac disease), tropical aphth, hypogammaglobulinemic aphtha, mucositis induced by chemotherapy or radiation therapy, diarrhea induced by chemotherapy or radiation therapy, low-grade inflammation , metabolic endotoxemia, necrotizing enterocolitis, primary biliary cirrhosis, hepatitis, fatty liver disease (including parental nutrition-related intestinal atrophy, parenteral nutrition-associated liver disease (PNALD), nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) or gastrointestinal side effects of inflammatory conditions such as pancreatitis or graft versus host disease (GVHD).

In some aspects, a pharmaceutical composition of the invention is administered to a human subject in need of prevention or treatment of obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, inadequate glucose control, glucose tolerance Sex, dyslipidemia (such as elevated LDL levels or low HDL/LDL ratio), diabetes (such as type 2 diabetes, gestational diabetes), prediabetes, metabolic syndrome, or high blood pressure.

In some aspects, the pharmaceutical composition of the present invention is administered to a human individual to promote a biological effect selected from the group consisting of: increasing intestinal mass, improving intestinal function (particularly intestinal barrier function) in an individual in need thereof , Increase intestinal blood flow, repair intestinal damage or dysfunction.

In some aspects, a medicament of the invention is administered to a human subject in need of preventing or treating intestinal dysfunction or injury caused by or associated with GVHD and preventing or treating side effects such as diarrhea caused by or associated with GVHD combination.

In some aspects, a pharmaceutical composition of the invention is administered to a human subject in need of prevention or treatment of obesity, morbid obesity, obesity-related gallbladder disease, and obesity-induced sleep apnea.

In some aspects, a pharmaceutical composition of the invention is administered to a human subject in need of improved glucose tolerance and/or glucose control. In some aspects, a pharmaceutical composition of the invention is administered to a human subject in need of modulating (eg, increasing) circulating cholesterol levels, capable of lowering circulating triglyceride or LDL levels, and increasing HDL/LDL ratio.

Administration In some aspects, the pharmaceutical compositions of the invention are aqueous compositions. In some aspects, the pharmaceutical compositions of the present invention are suitable for administration by means of a syringe (optionally a pen-like syringe) subcutaneously ( s.c. ) , intramuscularly ( i.m. ) or intravenously ( i.v. ) Injection is for parenteral administration. In some aspects, the pharmaceutical compositions of the invention are suitable for subcutaneous injection into human subjects. In some aspects, the pharmaceutical compositions of the invention are suitable for intravenous injection into human subjects.

In some aspects, isotonic parenteral pharmaceutical compositions of the invention are suitable for single dose administration. In some aspects, isotonic parenteral pharmaceutical compositions for injection are adapted for injection in a single-use device. In some aspects, the single use device is selected from an injection pen or a single use syringe. In some aspects, the isotonic parenteral pharmaceutical compositions of the invention are suitable for multiple dose administration.

In some aspects, the isotonic parenteral pharmaceutical compositions of the invention are administered to a human subject in a volume that allows delivery to the subject of a total amount of the GLP-1/GLP-2 dual agonist of about 1 mg to about 25 mg. Administered by subcutaneous injection. In some aspects, the isotonic parenteral pharmaceutical compositions of the invention are administered subcutaneously to a human subject in a volume that allows delivery to the subject of a total amount of about 1 mg or greater of the GLP-1/GLP-2 dual agonist. Injection administration. In some aspects, the isotonic parenteral pharmaceutical compositions of the present invention are in a volume (preferably Human subjects are administered by subcutaneous injection in a volume that allows delivery of a total amount of about 6 mg to about 25 mg of the GLP-1/GLP-2 dual agonist to the subject. In some aspects, the isotonic parenteral pharmaceutical compositions of the invention are administered subcutaneously to a human subject in a volume that allows delivery to the subject of a total amount of about 1 mg or greater of the GLP-1/GLP-2 dual agonist. Injection administration. In some aspects, an isotonic parenteral pharmaceutical composition of the invention is administered subcutaneously to a human subject in a volume that permits delivery to the subject of a total amount of about 2 mg or greater of the GLP-1/GLP-2 dual agonist. Injection administration. In some aspects, an isotonic parenteral pharmaceutical composition of the invention is administered subcutaneously to a human subject in a volume that permits delivery to the subject of a total amount of about 3 mg or greater of the GLP-1/GLP-2 dual agonist. Injection administration. In some aspects, an isotonic parenteral pharmaceutical composition of the invention is administered subcutaneously to a human subject in a volume that permits delivery to the subject of a total amount of about 4 mg or greater of the GLP-1/GLP-2 dual agonist. Injection administration. In some aspects, an isotonic parenteral pharmaceutical composition of the invention is administered subcutaneously to a human subject in a volume that permits delivery to the subject of a total amount of about 5 mg or greater of the GLP-1/GLP-2 dual agonist. Injection administration. In some aspects, an isotonic parenteral pharmaceutical composition of the invention is administered subcutaneously to a human subject in a volume that permits delivery to the subject of a total amount of about 6 mg or greater of the GLP-1/GLP-2 dual agonist. Injection administration. In some aspects, an isotonic parenteral pharmaceutical composition of the invention is administered subcutaneously to a human subject in a volume that permits delivery to the subject of a total amount of about 9 mg or greater of the GLP-1/GLP-2 dual agonist. Injection administration. In some aspects, an isotonic parenteral pharmaceutical composition of the invention is administered subcutaneously to a human subject in a volume that permits delivery to the subject of a total amount of about 10 mg or greater of the GLP-1/GLP-2 dual agonist. Injection administration. In some aspects, the isotonic parenteral pharmaceutical composition of the invention is such that a total amount of about 7, 8, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 A volume of , 21, 22, 23, 24 or 25 mg or greater of the GLP-1/GLP-2 dual agonist is administered to a human subject by subcutaneous injection.

Biological Activity In some aspects, the peptide comprised in the pharmaceutical composition of the invention is a peptide according to Formula A and SEQ ID NO: 1, which has been previously described in patent application WO2018104561, which describes the Compounds, their preparation and purification and biological activity (Table 5, WO2018104561). Example 2 in WO2018104561 includes data on in vitro potency at the GLP-1 and GLP-2 receptors.

Chemical Stability: Purity Compositions of the invention, such as the isotonic parenteral pharmaceutical composition of the invention, provide good chemical stability. In other words, in the compositions of the invention, the GLP-1/GLP-2 dual agonist remains stable during storage. The chemical stability may be comparable with respect to an equivalent composition not comprising a preservative according to the invention as described herein.

In some aspects, the compositions of the invention have good relative purity. The relative purity may be comparable or improved relative to an equivalent composition not comprising a preservative according to the invention as described herein.

When referring herein to the "chemical stability" of a composition of the invention, this means the chemical stability of the GLP-1/GLP-2 dual agonist contained in the composition. In some aspects, the chemical stability of a GLP-1/GLP-2 dual agonist is determined using Assay I described herein.

The chemical stability of the GLP-1/GLP-2 dual agonist at time point Y in any of the test compositions disclosed herein can be expressed as GLP-1/GLP-2 dual agonist The relative purity, X ^Y , was determined by measuring the absolute purity X' of the GLP-1/GLP-2 dual agonist, and with the GLP-1/GLP-2 dual agonist at day zero (Day 0 ) is normalized to the absolute purity X ⁰ , wherein the absolute purities are determined by HPLC at a given time point Y by identifying the purity of the peak corresponding to the GLP-1/GLP-2 dual agonist.

Thus, at day zero (Day 0), the absolute purity X' is the same as the absolute purity ^X0 and therefore the chemical stability of the GLP-1/GLP-2 dual agonist in the test composition, expressed as the relative purity ^XY =100%, where Y=day 0.

Relative purity can be calculated as follows: X ^Y = (X ' /X ⁰ )*100 where X is the relative purity at a given time point Y, X ⁰ is the absolute purity on day 0 and X' is the given time point The absolute purity of Y, wherein the absolute purity of the GLP-1/GLP-2 dual agonist in the test composition X ⁰ or X' is determined by HPLC to identify the corresponding GLP-1/GLP-2 dual agonist peak purity.

It was surprisingly found that high compared to low peptide concentrations had no or little effect on the degradation (chemical stability) of the peptides in the presence of the preservatives m-cresol and phenol.

The relative purity at a given time point can be calculated by multiplying the slope of the purity by the number of weeks of storage and subtracting the modulus of this value from 100%.

In some aspects, the one or more GLP-1/GLP-2 dual agonists, such as CPD1 or any pharmaceutically acceptable pharmaceutical composition thereof, is stored, for example, at 40° C. for at least 2 weeks. The relative purity of the salt is at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some aspects, the one or more dual GLP-1/GLP-2 agonists, such as CPD1 or any pharmaceutically acceptable agonist thereof, are stored, e.g., at least 25°C, for at least 26 weeks in the pharmaceutical composition of the invention. Accepted salts have a relative purity of at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%. In some aspects, the one or more GLP-1/GLP-2 dual agonists, such as CPD1 or any pharmaceutically acceptable agonist thereof, are stored, e.g., at least 5° C., for at least 52 weeks in the pharmaceutical composition of the invention. Accepted salts have a relative purity of at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.

Chemical Stability: Oligomerization Peptides in solution can aggregate to form therapeutically inactive covalently linked oligomers. Peptide oligomerization can be measured using size exclusion chromatography (SEC) such as described herein in Assay II.

Various analytical techniques for measuring peptide stability are available in the art and are reviewed, for example, in Peptide and Protein Drug Delivery , 247-301, Ed. Vincent Lee, Marcel Dekker, Inc., New York, NY, Pubs. (1991 ) and Jones, A. Adv . Drug Delivery Rev. 10: 29-90 (1993) .

It was surprisingly found that, in the presence of the preservatives m-cresol and phenol, high peptide concentrations reduce the amount of covalent oligomers compared to low peptide concentrations and there is a plateau effect at peptide concentrations above 6 mg/mL (plateau effect).

The relative total non-oligomerized peptide at a given time point can be calculated by multiplying the oligomer slope by the number of weeks of storage and subtracting the modulus of this value from 100%.

In some aspects, the one or more GLP-1/GLP-2 dual agonists, such as CPD1 or any pharmaceutically acceptable pharmaceutical composition thereof, is stored, for example, at 40° C. for at least 2 weeks. The relative total non-oligomerized peptide (monomer) of the salt is at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some aspects, the one or more GLP-1/GLP-2 dual agonists, such as CPD1 or any pharmaceutically acceptable pharmaceutical composition thereof, are stored, e.g. The relative total non-oligomerized peptide of the salt is at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% %, 98% or 99%. In some aspects, the one or more GLP-1/GLP-2 dual agonists, such as CPD1 or any pharmaceutically acceptable pharmaceutical composition thereof, are stored, for example, at 5° C. for at least 52 weeks after storage of the pharmaceutical composition of the invention The relative total non-oligomerized peptide (monomer) of the salt is at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. list of abbreviations abbreviation Explanation iv Intravenous sc Subcutaneous HPLC HPLC ND Not determined NP Particle free part/cont Particles per container RP-HPLC reverse phase high performance liquid chromatography CPD compound SEQ ID NO serial identification number

Terms and Definitions When terms such as " about " and " approximately " are used with respect to numerical values, those skilled in the art will immediately recognize that any effect or result that may be associated with a given value may be obtained within a certain tolerance of the specified value. As used herein, the term " about " therefore means within a reasonable range of the stated value, such as plus or minus 10%. When the term " about " is used with respect to chemical stability in this patent application, the reasonable vicinity is less than 2%, such as 0.5% or 0.75%, 1% or 1.5%.

As used herein, the term " solvent " is intended to be a substance that dissolves a solute (a chemically distinct liquid, solid or gas) to produce a solution. Solvents are usually liquids, but can also be solids, gases or supercritical fluids. Solvents are generally classified by polarity, and are considered polar or non-polar, as indicated by the dielectric constant. In general, solvents with a dielectric constant greater than about 5 are considered " polar " and solvents with a dielectric constant less than 5 are considered " non-polar ".

Non-limiting Aspects of the Invention The following portion of this specification contains certain non-limiting aspects of the invention. The aspects described below may be combined with any of the aspects of the invention described above and below and herein. 1. A composition comprising: (a) one or more GLP-1/GLP-2 dual agonists comprising general formula A: H[Aib]EG-X5-F-X7-SELATILD-[Ψ]- QAARDFIAWLI-X28-HKITD (A), where X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and at least one of X5 and X7 or T, wherein [Ψ] refers to the L or D lysine residue, wherein the albumin binding moiety binds to the GLP-1/GLP-2 dual agonist, and wherein the albumin binding moiety is [K( [17-carboxy-heptadecyl]-isoGlu)]; (b) one or more preservatives, wherein the one or more preservatives comprise or be m-cresol and/or phenol; and (c) phosphate buffer liquid. 2. The composition according to aspect 1, wherein the composition is an isotonic parenteral pharmaceutical composition. 3. The composition of any of the preceding aspects, wherein the one or more preservatives comprise or are m-cresol, preferably wherein the m-cresol is present at a concentration of about 1.15 mg/mL to about 5.15 mg/mL , more preferably wherein the m-cresol is present at a concentration of about 3.15 mg/mL. 4. The composition of any of the preceding aspects, wherein the one or more preservatives comprise or are phenol, preferably wherein the phenol is present at a concentration of from about 2.5 mg/mL to about 8.5 mg/mL, more preferably wherein The phenol was present at a concentration of about 5.5 mg/mL. 5. The composition of any one of the preceding aspects, wherein the phosphate buffer is present at a concentration of about 5 mM to about 50 mM, preferably wherein the phosphate buffer is present at a concentration of about 20 mM. 6. The composition according to any one of the preceding aspects, wherein the phosphate buffer is a sodium phosphate buffer, preferably wherein the sodium phosphate buffer is selected from disodium phosphate or sodium dihydrogen phosphate, or a combination thereof. 7. The composition according to any one of the preceding aspects, wherein the pH of the composition is from about pH 6.0 to about pH 8.5, preferably from about pH 6.5 to about pH 8.5, preferably from about pH 7.0 to about pH 8.0 , a more optimal pH is about pH 8.0. 8. The composition of any one of the preceding aspects, wherein the one or more GLP-1/GLP-2 dual agonists have the general formula B: H[Aib]EG-X5-FT-SELATILD-[Ψ] -QAARDFIAWLI-X28-HKITD (B), wherein X5 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and wherein [Ψ] refers to L or D lysine residue, wherein the albumin binding moiety binds to the GLP-1/GLP-2 dual agonist, and wherein the albumin binding moiety is [K([17-carboxy-heptadecyl]-isoGlu)] . 9. The composition of any one of the preceding aspects, wherein the one or more GLP-1/GLP-2 dual agonists comprise the following sequence: H[Aib]EGSFTSELATILD[Ψ]QAARDFIAWLIQHKITD (SEQ ID NO: 1) . 10. The composition according to any one of the preceding aspects, wherein the one or more GLP-1/GLP-2 dual agonists are: Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecyl or Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecyl]-isoGlu)]QAARDFIAWLIQHKITD-NH ₂ (CPD1NH ₂ ), or A pharmaceutically acceptable salt of CPD1OH or CPD1NH ₂ , preferably a chloride salt of CPD1OH or CPD1NH ₂ . 11. The composition of any one of the preceding aspects, wherein the GLP-1/GLP-2 dual agonist is at a concentration of at least about 1 mg/mL, preferably about 1 mg/mL to about 33 mg/mL Such as about 2 mg/mL to about 33 mg/mL, such as about 1 mg/mL to about 25 mg/mL, such as about 6 mg/mL to about 25 mg/mL. 12. The composition of aspect 11, wherein the GLP-1/GLP-2 dual agonist is present at a concentration of about 2 mg/mL, about 15 mg/mL or about 25 mg/mL. 13. The composition according to any one of the preceding aspects, wherein the composition further comprises one or more tonicity agents. 14. The composition according to aspect 13, wherein the one or more tonicity agents comprise or are mannitol, preferably D-mannitol. 15. The composition according to aspect 14, wherein the mannitol is in the range of about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM concentration exists. 16. The composition of aspect 13, wherein the one or more tonicity agents comprise or are NaCl, preferably wherein the NaCl is present at about 50 mM to about 450 mM, preferably about 65 mM to about 165 mM, preferably about 125 mM Present in mM concentrations. 17. The composition of any one of the preceding aspects, wherein the composition has an osmolality of from about 230 mOsmol/kg to about 370 mOsmol/kg. 18. The composition according to any one of the preceding aspects, wherein the composition further comprises a solvent, preferably water. 19. The composition according to any one of the preceding aspects, wherein the one or more preservatives are cresol at a concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, wherein the The concentration of phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM, wherein the composition further comprises a concentration of about 130 mM to about 330 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM of mannitol, preferably D-mannitol, and wherein the pH of the composition is from about pH 7.0 to about pH 8.0, preferably about pH 8.0. 20. The composition of any one of the preceding aspects, wherein the one or more preservatives are phenol at a concentration of about 2.5 mg/mL to about 8.5 mg/mL, preferably about 5.5 mg/mL, wherein the phosphate The concentration of the buffer is about 5 mM to about 50 mM, preferably about 20 mM, wherein the composition further comprises about 130 mM to about 330 mM, preferably about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM of mannitol, preferably D-mannitol, and wherein the pH of the composition is from about pH 6.0 to about pH 8.5, preferably from about pH 7.0 to about pH 8.0, more preferably Optimal pH 8.0. 21. The composition according to any one of the preceding aspects, wherein the one or more GLP-1/GLP-2 dual agonists are CPD1OH or CPD1NH ₂ , preferably CPD1OH, or a chloride salt of CPD1OH or CPD1NH ₂ , more preferably Preferably the chloride salt of CPD1OH, wherein the one or more preservatives are cresol at a concentration of about 1.15 mg/mL to about 5.15 mg/mL, preferably about 3.15 mg/mL, wherein the concentration of the phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM, and wherein the composition further comprises mannitol (preferably D-mannitol) at a concentration of about 230 mM, water and for adjusting the pH to a pH of about Sodium hydroxide and/or hydrochloric acid at pH 8.0. 22. The composition according to any one of the preceding aspects, wherein the one or more GLP-1/GLP-2 dual agonists are CPD1OH or CPD1NH ₂ , preferably CPD1OH, or a chloride salt of CPD1OH or CPD1NH ₂ , more preferably Preferably the chloride salt of CPD1OH, wherein the one or more preservatives are phenol at a concentration of about 2.5 mg/mL to about 8.5 mg/mL, preferably about 5.5 mg/mL, wherein the concentration of the phosphate buffer is about 5 mM to about 50 mM, preferably about 20 mM, and wherein the composition further comprises mannitol (preferably D-mannitol) at a concentration of about 230 mM, water and a pH for adjusting the pH to about pH 8.0 sodium hydroxide and/or hydrochloric acid. 23. The composition according to any one of the preceding aspects, wherein the composition is suitable for subcutaneous ( s.c. ) or intravenous ( i.v. ) injection into a human subject . 24. The composition according to any one of the preceding aspects, which is used for: (i) increasing intestinal mass, improving intestinal function, increasing intestinal blood flow or repairing intestinal damage or dysfunction in individuals in need; or (ii) prevention or treatment of malabsorption, ulcers, short bowel syndrome, blind tube syndrome, inflammatory bowel disease, irritable bowel syndrome, pouchitis, celiac disease, tropical aphth, hypogammaglobulinemia in individuals in need Aphtha, mucositis induced by chemotherapy or radiation therapy, diarrhea induced by chemotherapy or radiation therapy, low-grade inflammation, metabolic endotoxemia, necrotizing enterocolitis, primary biliary cirrhosis, hepatitis, Gastrointestinal side effects of fatty liver disease or inflammatory conditions; or (iii) reducing or inhibiting weight gain, reducing gastric emptying or intestinal transit, reducing food intake, reducing appetite, or promoting weight loss in individuals in need thereof; or (iv) preventing Or to treat obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, inadequate glucose control, glucose tolerance, dyslipidemia, diabetes, prediabetes, metabolic syndrome, or hypertension in an individual in need thereof. 25. A method for preserving a composition comprising one or more GLP-1/GLP-2 dual agonists comprising the general formula A: H[Aib]EG-X5-F-X7-SELATILD-[ Ψ]-QAARDFIAWLI-X28-HKITD (A), wherein X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and among X5 and X7 at least one of which is T, and wherein [Ψ] refers to the L or D lysine residue, wherein the albumin binding moiety binds to the GLP-1/GLP-2 dual agonist, and wherein the albumin binding moiety is [K([17-carboxy-heptadecyl]-isoGlu)]; wherein the composition comprises phosphate buffered saline; and wherein the method comprises adding one or more preservatives to the composition, wherein one or more of the preservatives comprise or be m-cresol and/or phenol. 26. Use of a preservative for preserving compositions comprising one or more GLP-1/GLP-2 dual agonists comprising the general formula A: H[Aib]EG-X5-F-X7- SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD (A), wherein X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and X5 and at least one of X7 is T, and wherein [Ψ] refers to an L or D lysine residue, wherein the albumin binding moiety binds to the GLP-1/GLP-2 dual agonist, and wherein the albumin binding moiety binds to the GLP-1/GLP-2 dual agonist, and wherein the albumin The protein binding moiety is [K([17-carboxy-heptadecyl]-isoGlu)]; wherein the composition comprises phosphate buffer; and wherein the preservative comprises or is m-cresol and/or phenol.

General method used Method I - Method for preparation of GLP-1/GLP-2 dual agonist GLP-1/GLP-2 dual agonist as described in Example 1 and according to General in patent application WO2018/104561 Peptide Synthesis synthesis.

CPD1 (corresponding to compound 18 in WO2018/104561) was synthesized using solid-phase peptide synthesis (SPPS) method and standard Fmoc coupling method. After completion of the synthesis, the peptide sequence was deprotected and cleaved from the solid support, and the crude peptide was purified using preparative reverse phase HPLC. Conversion of the peptide to an acceptable salt form (HCl, acetate or Na) and lyophilization yields the final CPD1 drug substance.

Method II - Preparation and Analytical Methods of Pharmaceutical Compositions Sample solutions for laboratory scale GLP-1/GLP-2 dual agonist drug substance (CPD1) were prepared according to method I and dissolved in MilliQ water (MQW), A stock solution of 40 mg/mL active pharmaceutical ingredient (API) was obtained. Measure the pH. This was followed by addition and mixing of ingredients as specified in Tables 5-8, and then adjusting the pH using 1 M NaOH/HCl as necessary to achieve the proper pH. Final concentrations of CPD1 were 2, 4, 6, 8, 10, 15 or 25 mg/mL as indicated in the Tables and Examples in this application. Laboratory scale compositions are prepared in volumes between 0.5 and about 2 mL.

For stability testing, samples were stored in a dark room (ie with lights off) at 25°C for 26 or 52 weeks, or at 40°C for 2 weeks (as indicated in the Examples). Samples were analyzed by RP-HPLC and SEC-HPLC according to assays I and II, respectively.

Analysis I - Measurement of GLP-1/GLP-2 dual agonist purity using RP-HPLC and determination of standardized GLP-1/GLP-2 dual agonist purity in % GLP-1/GLP-2 dual agonist The chemical stability of an agonist (peptide) is expressed herein as the relative purity of the peptide peak (i.e. the major peptide peak) as determined by HPLC at a given time point, and is expressed as the peptide peak at day zero (T=0). (ie the major peptide peak) was normalized for absolute purity, which was set to 100% normalized purity. The RP-HPLC method is capable of detecting CPD1 degradation products (deamidation, isomerization, hydrolysis and racemization). The RP-HPLC method cannot detect covalent oligomers, where two or more CPDl molecules are linked together via covalent chemical bonds - see Analysis II for more information on detecting covalent oligomers.

The chemical stability of the GLP-1/GLP-2 dual agonist (peptide) prepared according to method I contained in the parenteral pharmaceutical composition as prepared according to method II was analyzed according to the following method:

Analysis was performed using a Dionex Ultimate 3000 HPLC system (Thermo Fisher) giving a linear gradient at a flow rate of approximately 0.5 mL/min. The mobile phase composition consisted of 0.3% trifluoroacetic acid (TFA) in 90% acetonitrile/10% MQW and 0.3% TFA in MQW. Detection was performed using a wavelength of 215 nm. The injection volume was 2 μg of peptide. The column used for HPLC analysis is Phenomenex Kinetex C18, 150, particle size 3.0 mm, 2.6 µm. Run time is 25 minutes. Table 4 - RP-HPLC method details method file name LP401.073.02 String Phenomenex Kinetex C18, 150, 3.0 mm, 2.6 µm gradient(time; B%) 0;40, 20;70, 20.01;95, 22;95, 22.01;40, 25;40 Solvent A 0.3% TFA in MQW Solvent B 0.3% TFA in MeCN flow rate 0.500 mL/min Injection volume 2 µg Column temperature 25°C Autosampler temperature 4°C UV detection 215 nm

Results are presented in Tables 5-8 as degradation slopes calculated from normalized purity results. The slope is a measure of how rapidly CPD1 degrades. Lower values (ie otherwise from 0) indicate higher degradation.

Analysis II - Method for Evaluation of Covalent Oligomers by Size Exclusion Chromatography (SEC) Size exclusion layer was performed on a Dionex Ultimate 3000 HPLC system (Thermo Fisher) using isocratic elution at a flow rate of 0.5 mL/min Analytical method (SEC) experiment. The mobile phase consisted of 0.1% TFA, 45% acetonitrile/MQW. Detection was performed using a wavelength of 215 nm. The injection volume was 2 μg of peptide. The column used for SEC analysis was TSKgel SuperSW2000 (TOSOH Company), 4 µm, 30×4.6 mm, and the column temperature was 25°C. Run time is 12 minutes.

The SEC method is capable of detecting covalent oligomers, in which two or more CPD1 molecules are linked together via covalent chemical bonds.

Oligomerization data are presented in Tables 5-8. Data are presented as slopes calculated from covalent oligomer results. The slope is a measure of how rapidly CPD1 forms covalent oligomers. Higher numbers indicate higher covalent oligomer formation.

Examples These examples investigate the chemical stability and oligomerization of CPD1 in compositions containing different preservatives and buffers stored at different temperatures for different lengths of time.

CPD1 was prepared according to Method I. Pharmaceutical compositions (ie formulations) comprising different preservatives were prepared and stored according to Method II. The peptide is CPD1OH, which comprises the amino acid sequence of formula A. _CPD1OH is interchangeable with CPD1NH2.

The chemical stability of CPD1 is expressed as the slope of the relative purity of the peptide over time. The slope was determined by measuring the absolute purity of the peptide peak (i.e. the main CPD1 peak) at each time point using HPLC as described in Analysis 1, and then comparing this value to the absolute purity of the peptide peak at T=0 (which was set as 100% purity) normalized to obtain the relative purity percentage at each time point to determine. The slope over time is calculated from these relative purity values.

Oligomerization of CPD1 is represented as the slope of the proportion of covalent oligomers over time, as determined using Assay II at each time point.

Example 1 - Chemical stability and oligomerization of CPD1 in compositions comprising m-cresol or phenol In this example, no preservatives m-cresol or phenol were included and combinations containing 2 mg/mL or 10 mg/mL peptide The formulations (Formulations A-F) were stored at 25°C for 52 weeks or at 5°C for 52 weeks. The compositions of Formulations A-F are shown in Table 5 together with the chemical stability (purity) slope and oligomerization slope for each storage temperature and time period.

The total proportion of chemically stable non-oligomerized CPD1 in each formulation after storage can be calculated by subtracting the percentage of oligomerized peptide from the final percentage purity of the peptide. These results are shown in Table 6.

Example 2 - Chemical stability and oligomerization of CPD1 in compositions containing m-cresol or phenol stored at 40°C for 2 weeks The composition of phosphate buffer was stored at 40 °C for 2 weeks. The chemical stability (purity) slope and oligomerization slope for each formulation are shown in Table 7.

Example 3 - Chemical stability and oligomerization of CPD1 in compositions comprising m-cresol or phenol and varying peptide concentrations In this example, compositions comprising m-cresol or phenol preservatives and varying concentrations of CDP1 peptide Store at ℃ for 2 weeks. The chemical stability (purity) slope and oligomerization slope for each formulation are shown in Table 8.

The data show that in the presence of the preservatives m-cresol and phenol, high peptide concentrations reduce the amount of covalent oligomers compared to low peptide concentrations (see Table 7 for comparison of 2 mg/mL peptide with 25 mg/mL peptide ), and there was a plateau effect at concentrations above 6 mg/mL (see Table 8, showing oligomerization of peptides in the concentration range from 2 mg/mL to 25 mg/mL). In general, RP-HPLC purity data showed that high peptide concentrations had little or no effect on peptide degradation.

All publications mentioned in the above specification are incorporated herein by reference. Various modifications and variations of the described methods and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in biochemistry, molecular biology or related fields are intended to be within the scope of the following claims. Table 5 - CPD1 Purity and Oligomerization Slope of Formulations AF Formulation number Peptide concentration (mg/mL) pH Phosphate (mM) Mannitol (mM) m-cresol (mg/mL) Phenol (mg/mL) Purity slope 25 ℃ 52 weeks Purity slope 5 ℃ 52 weeks Oligomer slope 25 ℃ 52 weeks Oligomer slope 5 ℃ 52 weeks A 10 8 20 230 - - -0.21 -0.000 0.05 0.002 B 10 8 20 230 3.15 - -0.24 -0.004 0.08 0.003 C 10 8 20 230 - 5.5 -0.16* -0.006 0.03 0.004 D. 2 8 20 230 - - -0.18 -0.002 0.02 0.003 E. 2 8 20 230 3.15 - -0.31 -0.008 0.19 0.011 f 2 8 20 230 - 5.5 -0.22* -0.011 0.22 0.017 *Slope based on 13 weeks Table 6 - Percentage of chemically stable non-oligomerized CPD1 for Formulations AF Formulation number Main peak purity (%) after 52 weeks at 2 - 8 °C Oligomers (%) after 52 weeks at 2 - 8 °C Total chemically stable non-oligomerized CPD1 after 52 weeks at 2 - 8 °C (%) A 99.7 0.9 98.8 B 99.7 1.0 98.7 C 99.6 0.8 98.8 D. 99.5 0.9 98.6 E. 99.5 2.4 97.1 f 99.3 1.6 97.7 Table 7 - CPD1 Purity and Oligomerization Slope of Formulations 1-17 Formulation number Peptide concentration (mg/mL) pH Phosphate (mM) Mannitol (mM) m-cresol (mg/mL) Phenol (mg/mL) Normalized purity slope [%/ week ] 2 weeks, 40 ℃ Oligomer slope [%/ week ] 2 weeks, 40 ℃ 1 2 8 20 230 3.15 - -2.3 1.5 2 2 8 5 230 1.15 - -1.3 0.5 3 25 8 5 230 1.15 - -1.6 0.2 4 2 8 50 230 1.15 - -1.9 0.6 5 25 8 50 230 1.15 - -1.8 0.1 6 2 8 5 230 5.15 - -1.7 1.0 7 25 8 5 230 5.15 - -1.7 0.4 8 2 8 50 230 5.15 - -2.5 1.5 9 25 8 50 230 5.15 - -1.8 0.2 10 2 8 5 230 - 2.5 1.6 0.4 11 25 8 5 230 - 2.5 -1.4 0.2 12 2 8 50 230 - 2.5 -1.9 0.5 13 25 8 50 230 - 2.5 -1.7 0.1 14 2 8 5 230 - 8.5 -1.7 0.8 15 25 8 5 230 - 8.5 -1.5 0.2 16 2 8 50 230 - 8.5 -2.2 1.0 17 25 8 50 230 - 8.5 -1.7 0.2 Table 8 - CPD1 Purity and Oligomerization Slope (Different Peptide Concentrations) of Formulations 18-31 Formulation number Peptide concentration (mg/mL) pH Phosphate (mM) Mannitol ( mM) m-cresol (mg/mL) Normalized purity slope [%/ week ] 26/52 weeks, 25 ℃ * Oligomer slope [%/ week ] 26/52 weeks, 25 ℃ * 18 2 8 20 230 3.15 -0.3 0.19 19 4 8 20 230 3.15 -0.3 0.12 20 6 8 20 230 3.15 -0.2 0.08 twenty one 8 8 20 230 3.15 -0.3 0.09 twenty two 10 8 20 230 3.15 -0.2 0.08 twenty three 15 8 20 230 3.15 -0.3 0.07 twenty four 25 8 20 230 3.15 -0.3 0.07 25 2 8 20 230 - -0.2 0.02 26 4 8 20 230 - -0.2 0.04 27 6 8 20 230 - -0.2 0.03 28 8 8 20 230 - -0.2 0.04 29 10 8 20 230 - -0.2 0.05 30 15 8 20 230 - -0.3 0.03 31 25 8 20 230 - -0.3 0.03 *Slopes for 2-10 mg/mL formulations were calculated based on 52-week data, and 15-25 mg/mL sample slopes were based on 26-week data.

         
          <![CDATA[<110> Zealand Pharma A/S]]>
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          His Xaa Glu Gly Xaa Phe Thr Ser Glu Leu Ala Thr Ile Leu Asp Xaa
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                      20 25 30
          Asp
          
      

Claims (15)

A composition comprising: (a) one or more GLP-1/GLP-2 dual agonists comprising the general formula A: H[Aib]EG-X5-F-X7-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD (A), Wherein X5 is T or S; X7 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and at least one of X5 and X7 is T, wherein [Ψ] refers to the L or D lysine residue wherein the albumin binding moiety binds to the GLP-1/GLP-2 dual agonist, and Wherein the albumin binding moiety is [K([17-carboxy-heptadecyl]-isoGlu)]; (b) one or more preservatives, wherein the one or more preservatives comprise or are m-cresol and/or phenol; and (c) Phosphate buffer. The composition according to claim 1, wherein the composition is an isotonic parenteral pharmaceutical composition. The composition of any one of the preceding claims, wherein the one or more preservatives comprise or are m-cresol, preferably wherein the m-cresol is at a concentration of about 1.15 mg/mL to about 5.15 mg/mL present, more preferably wherein the m-cresol is present at a concentration of about 3.15 mg/mL. A composition according to any one of the preceding claims, wherein the one or more preservatives comprise or are phenol, preferably wherein the phenol is present at a concentration of from about 2.5 mg/mL to about 8.5 mg/mL, more preferably wherein the Phenol was present at a concentration of approximately 5.5 mg/mL. The composition of any one of the preceding claims, wherein the phosphate buffer is present at a concentration of about 5 mM to about 50 mM, preferably wherein the phosphate buffer is present at a concentration of about 20 mM. The composition according to any one of the preceding claims, wherein the phosphate buffer is a sodium phosphate buffer, preferably wherein the sodium phosphate buffer is selected from disodium phosphate or sodium dihydrogen phosphate, or a combination thereof. The composition according to any one of the preceding claims, wherein the pH of the composition is from about pH 6.0 to about pH 8.5, preferably from about pH 6.5 to about pH 8.5, preferably from about pH 7.0 to about pH 8.0 , a more optimal pH is about pH 8.0. The composition of any one of the preceding claims, wherein the one or more GLP-1/GLP-2 dual agonists have the general formula B: H[Aib]EG-X5-FT-SELATILD-[Ψ]-QAARDFIAWLI-X28-HKITD (B), Where X5 is T or S; X28 is Q, E, A, H, Y, L, K, R or S, and wherein [Ψ] refers to the L or D lysine residue wherein the albumin binding moiety binds to the GLP-1/GLP-2 dual agonist, and Wherein the albumin binding moiety is [K([17-carboxy-heptadecyl]-isoGlu)]. The composition of any one of the preceding claims, wherein the one or more GLP-1/GLP-2 dual agonists comprise the following sequence: H[Aib]EGSFTSELATILD[Ψ]QAARDFIAWLIQHKITD (SEQ ID NO: 1). A composition as in any one of the preceding claims, wherein the one or more GLP-1/GLP-2 dual agonists are: Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecyl] -isoGlu)]QAARDFIAWLIQHKITD-OH (CPD1OH); or Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecyl]-isoGlu)]QAARDFIAWLIQHKITD-NH ₂ (CPD1NH ₂ ), or CPD1OH or A pharmaceutically acceptable salt of CPD1NH ₂ , preferably a chloride salt of CPD1OH or CPD1NH ₂ . The composition of any one of the preceding claims, wherein the GLP-1/GLP-2 dual agonist is at a concentration of at least about 1 mg/mL, preferably at a concentration of about 1 mg/mL to about 33 mg/mL , such as at a concentration of about 2 mg/mL to about 33 mg/mL, such as at a concentration of about 1 mg/mL to about 25 mg/mL, such as at a concentration of about 6 mg/mL to about 25 mg/mL. The composition according to claim 11, wherein the GLP-1/GLP-2 dual agonist is present at a concentration of about 2 mg/mL, about 15 mg/mL or about 25 mg/mL. The composition according to any one of the preceding claims, wherein the composition further comprises one or more tonicity agents. The composition of claim 13, wherein the one or more tonicity agents comprise or are mannitol, such as D-mannitol, preferably wherein the mannitol is present at about 130 mM to about 330 mM, preferably It is present at a concentration of about 150 mM to about 300 mM, preferably about 190 mM to about 240 mM, preferably about 230 mM. The composition of claim 13, wherein the one or more tonicity agents comprise or are NaCl, preferably wherein the NaCl is from about 50 mM to about 450 mM, preferably from about 65 mM to about 165 mM, preferably about 125 mM concentration exists.